Figure 2. Glutaminase 1 (GLS1) is overexpressed in a mouse model of non-alcoholic steatohepatitis (NASH) of choline deficient and 0.1% methionine diet (0.1% MCDD)-fed rodents.

A. Hepatic Glutaminase 1 (GLS1), with higher magnification zoom shown in inset, and Glutaminase 2 (GLS2) immunostaining and respective quantifications. Scale bar corresponds to 100 μm. V-venous region; P-portal region; B. Hepatic GLS1 and GLS2 protein levels by Western blot analysis. Glyceraldehyde-3-phosphate (GAPDH) was used as a loading control; and C. Hepatic Gls1 and Gls2 mRNA levels in mice fed a choline deficient and 0.1% methionine diet (0.1% MCDD) against a standard chow diet (SC diet) during 2, 4 and 6 weeks. Data is shown as average ± SEM and Student’s t-test was used to compare groups. *p<0.05, **p<0.01 and ***p<0.001 are shown versus age- and gender-matched animals maintained on SC diet. D. Representative consecutive slides staining for GLS1, F4/80 and Sirius red staining in liver biopsies of animals maintained for four weeks or six weeks on 0.1% MCDD (fibrosis areas highlighted with white dashed line and GLS1 with yellow dashed line). E. Immunofluorescence double co-staining for GLS1 and albumin, a marker of hepatocytes, and alpha smooth muscle actin (aSMA), a marker of hepatic stellate cells (HSC) in 0.1% MCDD-fed mice for 4 weeks. (See also Supplemental Figure 1, Supplemental Figure 2, Supplemental Figure 3 and Supplemental Figure 4).